|
| Status |
Public on Oct 06, 2008 |
| Title |
Patient no. 6: treated |
| Sample type |
RNA |
| |
|
| Source name |
Liposarcoma culture from tumor 379, doxorubicin treated
|
| Organism |
Homo sapiens |
| Characteristics |
Sex: female
Age at operation: 58 years
Site: pelvis retroperitoneal
Size in cm: 4x7x9
Histological Subtype: myxoid/roundcell liposarcoma
Responder type: high
Grading: G3
Specimen character: metastasis
Previous radiation: no
Previous chemotherapy: no
|
| Treatment protocol |
24 h later, after having grown to a subconfluent layer, cell cultures were incubated with doxorubicin (0.5 ug/ml) for 24 h or an equal volume of PBS as control
|
| Growth protocol |
Primary human liposarcoma cultures were isolated by dissecting the tumor and digesting the minced samples enzymatically with 10 ml each of collagenase and dispase (10 mg/ml). The single cell suspension was depleted of red blood cells and cellular debris by centrifugation through a Ficoll-Hypaque density gradient. Liposarcioma cells were diluted and cultured during the whole experiment with Leibovitz's L-15 medium, supplemented with 2.0 mM glutamine and 10% fetal bovine serum in a humidified atmosphere in free air exchange with atmospheric air. Cells were seeded at a density of 2.00E+06 in 25 cm2 flasks.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Trizol extraction of total RNA was performed according to the manufacturer's instructions.
|
| Label |
biotin
|
| Label protocol |
Biotinylated cRNA were prepared according to the standard Affymetrix protocol from 5 ug total RNA (Expression Analysis Technical Manual, 2001, Affymetrix).
|
| |
|
| Hybridization protocol |
Following fragmentation, 10 ug of cRNA were hybridized for 16 hr at 45C on GeneChip Human Genome U133A Array. GeneChips were washed and stained in the Affymetrix Fluidics Station 400.
|
| Scan protocol |
GeneChips were scanned using the Hewlett-Packard GeneArray Scanner G2500A.
|
| Description |
Gene expression data from primary liposarcom cultures incubated with doxorubicin (0.5 µg/ml) for 24 h
|
| Data processing |
The data were analyzed with GCOS 1.4 using Affymetrix default analysis settings and global scaling as normalization method. The trimmed mean target intensity of each array was arbitrarily set to 1000.
|
| |
|
| Submission date |
Sep 29, 2008 |
| Last update date |
Jan 13, 2009 |
| Contact name |
Ludger Klein-Hitpass |
| E-mail(s) |
ludger.klein-hitpass@uni-essen.de
|
| Phone |
+49 201 723 85552
|
| Organization name |
Institut fuer Zellbiologie
|
| Department |
Universitaetsklinikum
|
| Lab |
BioChip Lab
|
| Street address |
Virchowstr. 173
|
| City |
Essen |
| ZIP/Postal code |
D-45122 |
| Country |
Germany |
| |
|
| Platform ID |
GPL96 |
| Series (1) |
| GSE12972 |
Heterogeneous in vitro effects of doxorubicin on gene expression in primary human liposarcoma cultures |
|
